CN105018355A - Breeding method for hypsizigus marmoreus strains - Google Patents
Breeding method for hypsizigus marmoreus strains Download PDFInfo
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- CN105018355A CN105018355A CN201510516667.1A CN201510516667A CN105018355A CN 105018355 A CN105018355 A CN 105018355A CN 201510516667 A CN201510516667 A CN 201510516667A CN 105018355 A CN105018355 A CN 105018355A
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Abstract
The invention discloses a breeding method for hypsizigus marmoreus strains. The breeding method comprises steps as follows: (1), a culture medium is prepared; (2), bottling and sterilizing are performed; (3), the culture medium is poured into culture vessels; (4), inoculation is performed: a black line is marked on a bottom cover of each culture vessel to divide the surface of the bottom cover into two parts equally, and hypsizigus marmoreus mother strains and trichoderma viride pathogenic bacteria are inoculated on two opposite points 3 cm away from the center of a flat plate respectively; (5), culturing is performed: the culture vessels are put in a dark and airtight environment at the temperature of 22 DEG C after inoculation for culturing, and the diameter of a bacterial colony in each culture vessel is recorded day by day; (6), after culturing, the antibacterial rate of the hypsizigus marmoreus strains to the trichoderma viride pathogenic bacteria in each culture vessel is calculated, and the hypsizigus marmoreus strains with a higher antibacterial rate in one culture vessel are selected as target strains. The hypsizigus marmoreus strains with higher trichoderma viride activity resistance can be cultured without mutagenesis with physical and chemical methods, the method is simple, convenient and easy to popularize, energy is seldom consumed, no pollution is caused to the environment, and the breeding method accords with the industrial policy about environment protection in China.
Description
Technical field
The invention belongs to the use production technical field of edible mushrooms, be specifically related to a kind of selection of Hypsizygus marmoreus bacterial classification.
Background technology
Viride be in the raw bacterium of domestic and international multiple wood and the raw bacteria cultivation of grass and bacterial classification production process time there is the most a kind of fungal disease.The Hypsizygus marmoreus speed of growth is comparatively slow, suppresses the ability of miscellaneous bacteria poor, and is subject to the impact of planting environment and cultivation condition, have certain inferior position compared with other factory edible fungi production kinds.Strain improvement is one of most critical technology in Hypsizygus marmoreus cultivation step, and be also just confined among a small circle owing to making Hypsizygus marmoreus cultivate by the restriction of technology and equipment condition, this is unfavorable for that the all-round popularization of Hypsizygus marmoreus industry is popularized.The seed selection of existing Hypsizygus marmoreus bacterial classification turns out the energetic Hypsizygus marmoreus bacterial classification of anti-viride mainly through the mutagenesis of physics and chemistry method, and this method complex process, input cost are high, are not easy to spread.
Summary of the invention
The technical problem to be solved in the present invention is to provide the selection of simple, the easy to operate Hypsizygus marmoreus bacterial classification of a kind of technique, and the method overcomes the problem that existing Hypsizygus marmoreus bacterial classification is subject to viride pathogen contamination in culturing process.
Technical scheme of the present invention comprises the following steps:
(1), substratum is prepared: fresh peeling potatoes will be cleaned, take that 180-220g is cut into small pieces, little bar or thin slice, add water 800-1200mL wherein, boil 20-30 min, removing slag with soaking the multilayer filtered through gauze wrung out in advance, collecting juice, and add glucose 18-22 g wherein, agar 18-22g stirs, and supply moisture and be settled to 1000ml, namely obtain substratum;
(2), bottling sterilizing: according to often bottled enter amount be the 55-65% of triangular flask volume, the substratum prepared is loaded triangular flask, and high-pressure sterilizing pot sterilizing is put in sealing;
(3), substratum pours culture dish into: after sterilizing completes, after waiting culture-liquid temp to cool, pour in multiple culture dish of sterilizing;
(4), inoculation: draw black line on each culture dish bottom area is halved, and inoculate Hypsizygus marmoreus mother kind and viride pathogenic bacteria respectively on 2 relative apart from plate center 2.5-3.5;
(5), cultivate: the culture dish inoculated all is put into the also airtight cultivation of temperature control 20-25 DEG C lucifuge in incubator, records the colony diameter of each culture dish day by day;
(6), cultivate after 4-6 days, calculate Hypsizygus marmoreus bacterial classification in each culture dish and, to the bacteriostasis rate of viride pathogenic bacteria, select the Hypsizygus marmoreus bacterial classification in the higher culture dish of bacteriostasis rate to be the target bacterial classification of institute's seed selection.
On such scheme basis, provide best technical scheme below, concrete steps are:
(1), substratum is prepared: fresh peeling potatoes will be cleaned, take that 200g is cut into small pieces, little bar or thin slice, add water 1000 mL wherein, boil 20-30 min, removing slag with soaking the 4 layers of filtered through gauze wrung out in advance, collecting juice, and add glucose 20 g wherein, agar 20g stirs, and supply moisture and be settled to 1000ml, namely obtain substratum;
(2), bottling sterilizing: according to often bottled enter amount be 60% of triangular flask volume, the substratum for preparing is loaded triangular flask, and high-pressure sterilizing pot sterilizing is put in sealing;
(3), substratum pours culture dish into: after sterilizing completes, after waiting culture-liquid temp to cool, pour in multiple culture dish of sterilizing;
(4), inoculation: draw black line on each culture dish bottom area is halved, and inoculate Hypsizygus marmoreus mother kind and viride pathogenic bacteria respectively on 2 relative apart from plate center 3cm;
(5), cultivate: the culture dish inoculated all is put into also temperature control 22 DEG C of airtight cultivations of lucifuge in incubator, records the colony diameter of each culture dish day by day;
(6), cultivate after 5 days, calculate Hypsizygus marmoreus bacterial classification in each culture dish and, to the bacteriostasis rate of viride pathogenic bacteria, select the Hypsizygus marmoreus bacterial classification in the higher culture dish of bacteriostasis rate to be the target bacterial classification of institute's seed selection.
Such scheme also can more preferably:
Triangular flask described in described step (2) is vial;
Autoclaving program in described step (3) is keep 30min at 121 DEG C.
The inoculation of described step (4) refers to and aseptically intercepts in Hypsizygus marmoreus and trichoderma viride block access substratum along colony edge with the punch tool of diameter 5mm respectively.
The airtight cultivation of lucifuge in described step (5) needs the culture dish by inoculation to cultivate at 22 DEG C.
Hypsizygus marmoreus in described step (6) deducts Hypsizygus marmoreus colony diameter again divided by the numerical value of the colony diameter gained of viride to the colony diameter that the bacteriostasis rate of pathogenic bacteria is viride.
Compared with prior art, the present invention has the following advantages:
1, selection of the present invention does not use the mutagenesis of physics and chemistry method just successfully can turn out the energetic Hypsizygus marmoreus bacterial classification of anti-viride, and this method is simple and convenient, be easy to promote.
2, the Hypsizygus marmoreus bacterial classification of method seed selection provided by the invention, the ability of opposing viride is strong, reduces because strain pollution inoculates the unnecessary loss caused.
3, method provided by the invention not electricity consumption, does not consume the energy, to environment without any pollution, meets the industry policy of national energy-saving environmental protection.
Embodiment
Embodiment 1
(1), substratum is prepared: fresh peeling potatoes will be cleaned, take that 200g is cut into small pieces, little bar or thin slice, add water 1000 mL wherein, boil 20-30 min, removing slag with soaking the 4 layers of filtered through gauze wrung out in advance, collecting juice, and add glucose 20 g wherein, agar 20g stirs, and supply moisture and be settled to 1000ml, namely obtain substratum;
(2), bottling sterilizing: according to often bottled enter amount be 60% of triangular flask volume, the substratum for preparing is loaded triangular flask, and high-pressure sterilizing pot sterilizing is put in sealing;
(3), substratum pours culture dish into: after sterilizing completes, after waiting culture-liquid temp to cool, pour in multiple culture dish of sterilizing;
(4), inoculation: draw black line by area 2 decile on each culture dish bottom, and inoculate Hypsizygus marmoreus mother kind and viride pathogenic bacteria respectively on 2 relative apart from plate center 3cm;
(5), cultivate: the culture dish inoculated all is put into also temperature control 22 DEG C of airtight cultivations of lucifuge in incubator, records the colony diameter of each culture dish day by day; The Hypsizygus marmoreus bacterial classification bacteriostasis rate that the 22 DEG C of lucifuge airtight cultivation seed selections of test proof obtain is the highest, compares 20 DEG C or 25 DEG C of average all height about 1 % of bacteriostasis rate.
(6), cultivate after 5 days, calculate Hypsizygus marmoreus bacterial classification in each culture dish and, to the bacteriostasis rate of viride pathogenic bacteria, select the Hypsizygus marmoreus bacterial classification in the higher culture dish of bacteriostasis rate to be the target bacterial classification of institute's seed selection; The bacterial classification bacteriostasis rate of the long or too short seed selection of incubation time is all not high, and optimum is 5 days, compares cultivation 4 days or 6 days average bacteriostasis rate height about 1%.
Respectively Hypsizygus marmoreus bacterial classification original before the Hypsizygus marmoreus bacterial classification of seed selection in embodiment and seed selection is done bacteriostasis rate test, test-results is in table one.
One, the Hypsizygus marmoreus bacterial classification bacteriostasis rate test of seed selection
Repeat embodiment 1 six times, seed selection obtains the energetic Hypsizygus marmoreus bacterial classification of No. 1-6 anti-viride respectively, and then tested respectively by the 1-6 Hypsizygus marmoreus bacterial classification of above-mentioned seed selection and determine its bacteriostasis rate, testing sequence is:
(a), choose the culture dish that above-mentioned steps (3) obtains, culture dish bottom is drawn black line by area 2 decile, and inoculate Hypsizygus marmoreus bacterial classification and the viride pathogenic bacteria of seed selection respectively on relative 2 of distance plate center 3cm;
(b), the culture dish inoculated is put in incubator and temperature control 22 DEG C of airtight cultivations of lucifuge, record the colony diameter of culture dish day by day;
(c), cultivated after, in calculating culture dish, Hypsizygus marmoreus bacterial classification is to the bacteriostasis rate of viride pathogenic bacteria, and Hypsizygus marmoreus deducts Hypsizygus marmoreus colony diameter again divided by the numerical value of the colony diameter gained of viride to the colony diameter that the bacteriostasis rate of pathogenic bacteria is viride.
Two, original before seed selection Hypsizygus marmoreus bacterial classification bacteriostasis rate test
Its bacteriostasis rate is determined in Hypsizygus marmoreus bacterial classification original before seed selection test, and testing sequence is:
(a), choose multiple culture dish that above-mentioned steps (3) obtains, each culture dish bottom is drawn black line by area 2 decile, and on two relative apart from plate center 3cm, inoculate original Hypsizygus marmoreus bacterial classification (namely Hypsizygus marmoreus mother plants) and viride pathogenic bacteria respectively;
(b), the culture dish inoculated is put into respectively in incubator and temperature control 22 DEG C of airtight cultivations of lucifuge, record the colony diameter of each culture dish day by day;
(c), cultivated after, to calculate in each culture dish Hypsizygus marmoreus bacterial classification to the bacteriostasis rate of viride pathogenic bacteria, and Hypsizygus marmoreus is to the bacteriostasis rate mean value of viride pathogenic bacteria in further each culture dish of calculating, Hypsizygus marmoreus deducts Hypsizygus marmoreus colony diameter again divided by the numerical value of the colony diameter gained of viride to the colony diameter that the bacteriostasis rate of pathogenic bacteria is viride.
D () repeating step (a)-(c) twice, obtains twice test Hypsizygus marmoreus respectively to the bacteriostasis rate mean value of viride pathogenic bacteria, namely contrast 1 and 2.
Table one
Hypsizygus marmoreus bacterial classification original before the Hypsizygus marmoreus bacterial classification of above-described embodiment seed selection and seed selection is done respectively the cultivation of two batches, cultivation result is as table two.
Table two
Contrast known by above table one and table two, the Hypsizygus marmoreus bacterial classification anti-viride vigor of seed selection of the present invention is stronger, bacteriostasis rate is than 4-6 percentage point high before seed selection, the activity of bacterial classification does not reduce simultaneously, output also improves a lot than contrast, Hypsizygus marmoreus single bottle of output reaches more than 230 grams, and growth cycle shortens 15 days, and fruiting improves more than 15%.The present invention is on the basis exceeding prior art index, and strain improvement is more convenient to operate, and investment is less, and energy consumption is lower, is obviously applicable to large-area popularization.
Above-described embodiment is only exemplary embodiments of the present invention, and those skilled in the art technical scheme according to the present invention can make simple adjustment completely, but identical with flesh and blood of the present invention, therefore fall into equally within protection scope of the present invention.
Claims (5)
1. a selection for Hypsizygus marmoreus bacterial classification, is characterized in that, comprises the following steps:
(1), substratum is prepared: fresh peeling potatoes will be cleaned, take that 180-220g is cut into small pieces, little bar or thin slice, add water 800-1200mL wherein, boil 20-30 min, removing slag with soaking the multilayer filtered through gauze wrung out in advance, collecting juice, and add glucose 18-22 g wherein, agar 18-22g stirs, and supply moisture and be settled to 1000ml, namely obtain substratum;
(2), bottling sterilizing: according to often bottled enter amount be the 55-65% of triangular flask volume, the substratum prepared is loaded triangular flask, and high-pressure sterilizing pot sterilizing is put in sealing;
(3), substratum pours culture dish into: after sterilizing completes, after waiting culture-liquid temp to cool, pour in multiple culture dish of sterilizing;
(4), inoculation: draw black line on each culture dish bottom area is halved, and inoculate Hypsizygus marmoreus mother kind and viride pathogenic bacteria respectively on 2 relative apart from plate center 2.5-3.5;
(5), cultivate: the culture dish inoculated all is put into the also airtight cultivation of temperature control 20-25 DEG C lucifuge in incubator, records the colony diameter of each culture dish day by day;
(6), cultivate after 4-6 days, calculate Hypsizygus marmoreus bacterial classification in each culture dish and, to the bacteriostasis rate of viride pathogenic bacteria, select the Hypsizygus marmoreus bacterial classification in the higher culture dish of bacteriostasis rate to be the target bacterial classification of institute's seed selection.
2. the selection of Hypsizygus marmoreus bacterial classification according to claim 1, is characterized in that, comprise the following steps:
(1), substratum is prepared: fresh peeling potatoes will be cleaned, take that 200g is cut into small pieces, little bar or thin slice, add water 1000 mL wherein, boil 20-30 min, removing slag with soaking the 4 layers of filtered through gauze wrung out in advance, collecting juice, and add glucose 20 g wherein, agar 20g stirs, and supply moisture and be settled to 1000ml, namely obtain substratum;
(2), bottling sterilizing: according to often bottled enter amount be 60% of triangular flask volume, the substratum for preparing is loaded triangular flask, and high-pressure sterilizing pot sterilizing is put in sealing;
(3), substratum pours culture dish into: after sterilizing completes, after waiting culture-liquid temp to cool, pour in multiple culture dish of sterilizing;
(4), inoculation: draw black line by area 2 decile on each culture dish bottom, and inoculate Hypsizygus marmoreus mother kind and viride pathogenic bacteria respectively on 2 relative apart from plate center 3cm;
(5), cultivate: the culture dish inoculated all is put into also temperature control 22 DEG C of airtight cultivations of lucifuge in incubator, records the colony diameter of each culture dish day by day;
(6), cultivate after 5 days, calculate Hypsizygus marmoreus bacterial classification in each culture dish and, to the bacteriostasis rate of viride pathogenic bacteria, select the Hypsizygus marmoreus bacterial classification in the higher culture dish of bacteriostasis rate to be the target bacterial classification of institute's seed selection.
3. the selection of Hypsizygus marmoreus bacterial classification according to claim 1 and 2, is characterized in that: the triangular flask described in described step (2) is vial.
4. the selection of Hypsizygus marmoreus bacterial classification according to claim 1 and 2, is characterized in that: the autoclaving program in described step (3) is keep 30min at 121 DEG C.
5. the selection of Hypsizygus marmoreus bacterial classification according to claim 1 and 2, it is characterized in that, the inoculation of described step (4) refers to and aseptically intercepts in Hypsizygus marmoreus and trichoderma viride block access substratum along colony edge with the punch tool of diameter 5mm respectively.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201510516667.1A CN105018355A (en) | 2015-08-21 | 2015-08-21 | Breeding method for hypsizigus marmoreus strains |
| CN201510677194.3A CN105154340B (en) | 2015-08-21 | 2015-10-18 | A kind of selection of true pleurotus cornucopiae strain |
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| CN201510516667.1A CN105018355A (en) | 2015-08-21 | 2015-08-21 | Breeding method for hypsizigus marmoreus strains |
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| CN201510677194.3A Active CN105154340B (en) | 2015-08-21 | 2015-10-18 | A kind of selection of true pleurotus cornucopiae strain |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109182450A (en) * | 2018-09-14 | 2019-01-11 | 江苏品品鲜生物科技有限公司 | A kind of selection of true pleurotus cornucopiae strain |
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| CN103004464B (en) * | 2012-11-12 | 2014-07-09 | 上海丰科生物科技股份有限公司 | Novel strain of hypsizigus marmoreus |
| CN103563653B (en) * | 2013-11-14 | 2015-05-20 | 江苏瑞光生物科技有限公司 | Method for bottle cultivation of pleurotus cornucopiae |
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| CN109182450A (en) * | 2018-09-14 | 2019-01-11 | 江苏品品鲜生物科技有限公司 | A kind of selection of true pleurotus cornucopiae strain |
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Application publication date: 20151104 |